Development of endometrium like 3D culture system using in vitro model implantation

Aim: For using both embryo and invasion models, we aimed to investigate three dimensional (3D) endometrium-like culture system invasive culture and epithelial mesenchymal transition mechanism. Via in vitro three-dimensional endometrium-like culture system, epithelial mesenchymal transition in both implantation and invasion process were analyzed using single or combine culture of endometrium epithelial (RL95-2) and trophoblast-like chiocarcinoma (JAR) cell lines on bacterial cellulose (BC), collagen foam (COL/K) and collagen fiber (COL/F) porous scaffolds. Materials and Methods: Three different types of tissue scaffolds were used in the study and RL95-2 and JAR cells were purchased commercially. Cells were cultured on appropriate media, after 80% confluency of the cells, they were plated onto scaffolds separately or combine. On culture time; 3., 5., 7., 10. and 14. days, the scaffolds were fixed with 4% paraformaldehyde and frozen sections were taken. Distributions of E-cadherin as an epithelial marker, N-cadherin, Vimentin, α-SMA and Syndecan-1 as a mesenchymal marker were analyzed by indirect immunoperoxidase or immunofluorescence staining methods. Characterization of scaffolds were analyzed by SEM method. Results: In the RL95-2 cells, immunoreactivities of E-cadherin, N-cadherin, Vimentin, α-SMA and Syndecan-1 were higher than JAR cells, in addition, E-cadherin and Syndecan-1 immunoreactivities in RL95-2 cells, N-cadherin and Syndecan-1 immunoreactivities in JAR cells were significantly higher. After the characterization of the scaffolds BC pore size of 97±28 μm, a COL/K pore size of 109±21 μm and a COL/F tissue scaffold of 0, 67 ± 0,14 µm in diameter were detected. The epithelial mesenchymal transition markers were stained in all three scaffolds, however, for epitelial mesenchymal transition BC and JAR cells, the mesenchymal properties of JAR cells on day 7, RL95-2 cells on day 14 on COL/K tissue scaffold were higher, whereas COL/F scaffold was supported epithelial properties of RL95-2 cells, in addition, immunoreactivities of E-cadherin, N-cadherin, Vimentin, α-SMA and Syndecan-1 were higher on combine culture of RL95-2 + JAR cells on this scaffold at 14. Day of the culture time. Conclusion: The newly developed 3D endometrium-like culture system using different scaffolds was supported the epithelial mesenchymal transition mechanisms in both cancer cell invasion and embryo implantation during different culture periods and it can be a preliminary study for further research.